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JP2014014366A Dna molecules and methods 有权
标题翻译： DNA分子和方法
公开(公告)号：JP2014014366A
公开(公告)日：2014-01-30
申请号：JP2013169290
申请日：2013-08-16
申请人： Morvus Technology Ltd , モーヴァステクノロジーリミテッド
发明人： NIGEL PETER MINTON , HEAP JOHN TIMOTHY
IPC分类号： C12N15/09 , C12N1/15 , C12N1/19 , C12N1/21 , C12N5/10
CPC分类号： C12N15/74 , C12N15/902
摘要： PROBLEM TO BE SOLVED: To provide DNA molecules and methods which allow for the efficient insertion into the genome of Clostridium spp. or other bacteria of the class Clostridia.SOLUTION: A DNA molecule comprises: a modified Group II intron (hereinafter abbreviated as intron II) which does not express the intron-encoded reverse transcriptase but which contains a modified selectable marker gene having an encoded region in the reverse orientation relative to the modified intron II, wherein a promoter is capable of causing expression of a selectable marker encoded by a single copy of the selectable marker gene in an amount sufficient for the selectable marker to alter the phenotype of a bacterial cell of the class Clostridia such that it can be distinguished; and a promoter for transcription of the modified intron II, the promoter being operably linked to the modified intron II; and wherein the modified selectable marker gene contains a Group I intron positioned in the forward orientation relative to the modified intron II so as to disrupt expression of the selectable marker gene, and wherein the DNA molecule allows for removal of the Group I intron from the RNA transcript of the modified intron II to leave a region encoding the selectable marker and allows for the insertion of the RNA transcript at a site in a DNA molecule in the bacterial cell.
摘要翻译：要解决的问题：提供允许有效插入梭菌属（Clostridium spp）基因组的DNA分子和方法。或其他类梭菌的细菌。解决方案：DNA分子包含：不表达内含子编码的逆转录酶但是含有编码区的修饰的选择标记基因的修饰的第II组内含子（以下简称为内含子II）相对于修饰的内含子II的相反方向，其中启动子能够引起由选择标记基因的单拷贝编码的选择性标记的表达量，其量足以使选择标记改变细胞细胞的表型可以区分梭菌，和修饰的内含子II的转录启动子，启动子可操作地连接到修饰的内含子II; 并且其中所述修饰的选择标记基因含有相对于修饰的内含子II位于正向取向的第I组内含子，以便破坏所述选择性标记基因的表达，并且其中所述DNA分子允许从所述RNA中去除所述第I组内含子修饰的内含子II的转录物离开编码可选择标记的区域，并允许将RNA转录物插入细菌细胞中的DNA分子中的位点。

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